This invention is directed to herpesvirus vaccines.
Herpesviruses are ubiquitous viruses which are the causative agents of numerous diseases in both humans and animals. These viruses are enveloped double stranded icosahedral DNA containing viruses, which envelope is acquired by budding of the nucleocapsid through the inner nuclear membrane. Members of the herpesvirus family which are important human pathogens include herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), varicella zoster virus (VZV), Epstein Barr virus (EBV), cytomegalovirus (CMV), and human herpesviruses type 6, type 7 and type 8 (HHV-6, HHV-7 and HHV-8).
The genome of HSV-1 encodes several glycoproteins which are important for viral pathogenesis. Four glycoproteins glycoprotein B (gB), glycoprotein D (gD), glycoprotein H (gH) and glycoprotein L (gL) are essential for virus infectivity in cells in culture and each appears to play a role in the mechanism by which the virus enters cells (Roop et al., 1993, J. Virol. 67:2285). Glycoprotein H is a 110 kDa protein encoded by the UL22 open reading frame of HSV-1 (Gompels and Minson, 1986, Virology 153:230). When gH is expressed in mammalian cell systems in the absence of other HSV-1 proteins it remains within the cell as an incompletely processed molecule (Foa-Tomasi et al., 1991, Virology 180:474; Roberts et al., 1991, Virology 184:609). When gH is expressed in cells which also express gL, gH and gL form a stable complex wherein fully processed gH is evident (Hutchinson et al., 1992, J. Virol. 66:2240). In addition, cells infected with a gL-negative mutant produce virus particles which lack both gH and gL (Roop et al., 1993, J. Virol. 67:2285). However, since transport of gH to the surface of cells is reported to occur in the absence of gL, gL may not be required in some systems for correct processing and transport of gH (Ghiasi et al., 1991, Virology 185:187).
A recombinant vaccinia virus expressing both gH and gL has been used to examine whether the gH-gL complex was capable of eliciting a protective immune response in mice. Weak levels of HSV-1 specific neutralizing antibody were evident in mice containing the complex and virus clearance from the site of challenge was only marginally enhanced when the gH-gL complex was administered to the animals compared with administration of gH alone (Browne et al., 1993, J. Gen. Virol. 74:2813).
Currently, there are no effective herpesvirus vaccines available for immunization of humans against any of the plethora of diseases caused by these pathogens, although subunit preparations comprising glycoprotein B, glycoprotein D, either alone or in combination are currently in clinical trials. Since herpesviruses cause recurrent and frequently fatal or permanently debilitating infections in humans and in other animals, there is a long felt need for such vaccines.
The invention relates to a vaccine comprising a soluble herpes simplex virus gHt-gL complex suspended in a pharmaceutically acceptable carrier.
In one aspect, the herpes simplex virus is selected from the group consisting of herpes simplex virus type 1 and herpes simplex virus type 2. Preferably, the herpes simplex virus is herpes simplex virus type 1.
In one embodiment, gHt is from a herpes simplex virus selected from herpes simplex virus type 1 and herpes simplex virus type 2.
In another embodiment, the gL is from a herpes simplex virus selected from herpes simplex virus type 1 and herpes simplex virus type 2.
In yet another embodiment, the gHt is from herpes simplex virus type 1 and comprises amino acid residues selected from the group consisting of 1-792, 1-648, 1-475, 1-324 and 1-275.
In a further embodiment, the gHt is herpes simplex virus type 1 gHt comprising amino acids 1-324.
In another embodiment, the gHt is herpes simplex virus type 1 gHt comprising amino acids 1-792 and the gL is herpes simplex virus type 1 gL comprising amino acids 1-168.
The vaccine of the invention may further include a substantially pure preparation of at least one of a herpes simplex virus gD, gB or gC.
Also included in the invention is a vaccine comprising an isolated DNA encoding a soluble herpes simplex virus gHt-gL complex suspended in a pharmaceutically acceptable carrier.
In one embodiment of this aspect of the invention, the vaccine includes an isolated DNA encoding at least one of a herpes simplex virus gD, gB or gC.
The invention also relates to a herpes simplex virus neutralizing antibody which specifically binds to a soluble herpes simplex virus gHt-gL complex.
The invention further relates to an isolated DNA encoding a herpes simplex virus neutralizing antibody which specifically binds to a soluble herpes simplex virus gHt-gL complex.
There is also included in the invention a method of immunizing a human patient against a herpes simplex virus infection comprising administering to the patient a vaccine comprising a soluble herpes simplex virus gHt-gL complex suspended in a pharmaceutically acceptable carrier.
The invention further relates to a method of immunizing a human patient against a herpes simplex virus infection comprising administering to the patient a vaccine comprising an isolated DNA encoding a soluble herpes simplex virus gHt-gL complex suspended in a pharmaceutically acceptable carrier.
Also included in the invention is a method of treating a herpes simplex virus infection in a human patient comprising administering to the patient a vaccine comprising a soluble herpes simplex virus gHt-gL complex suspended in a pharmaceutically acceptable carrier.
The invention further includes a method of treating a herpes simplex virus infection in a human patient comprising administering to the patient a vaccine comprising an isolated DNA encoding a soluble herpes simplex virus gHt-gL complex suspended in a pharmaceutically acceptable carrier.
In each of the aforementioned methods of immunizing a human patient or of treating a herpes simplex virus infection in a human patient, the methods should be construed to optionally include the administration of a substantially pure preparation of at least one of a herpes simplex virus gD, gB or gC, or the administration of an isolated DNA encoding at least one of a herpes simplex virus gD, gB or gC.
There is further provided a method of treating a herpes simplex virus infection in a human patient comprising administering to the patient a herpes simplex virus neutralizing antibody which specifically binds to a soluble herpes simplex virus gHt-gL complex wherein the antibody is suspended in a pharmaceutically acceptable carrier.
Also included is a method of treating a herpes simplex virus infection in a human patient comprising administering to the patient an isolated DNA encoding a herpes simplex virus neutralizing antibody which specifically binds to a soluble herpes simplex virus gHt-gL complex, wherein the DNA is suspended in a pharmaceutically acceptable carrier.
In addition, the invention relates to a preparation of a soluble herpes simplex virus gHt-gL complex and a substantially pure preparation of a soluble herpes simplex virus gHt-gL complex.
In a preferred embodiment, the herpes simplex virus is selected from the group consisting of herpes simplex virus type 1 and herpes simplex virus type 2. Preferably, the herpes simplex virus is herpes simplex virus type 1.
In one aspect, the complex is suspended in a pharmaceutically acceptable carrier.
In another aspect, the gHt comprises amino acid residues selected from the group consisting of 1-792, 1-648, 1-475, 1-324 and 1-275.
In yet another aspect, the gHt comprises amino acids 1-324.
In another aspect, the gHt comprises amino acids 1-792 and the gL comprises amino acids 1-168.
The invention further relates to a cell, the DNA of the cell encoding a soluble herpes simplex virus gHt-gL complex.
In one aspect, the cell is HL-7.
There is also provided in the invention a method of modifying a cell to render it capable of secreting a soluble herpes simplex virus gHt-gL complex comprising introducing into the cell DNA encoding a truncated form of herpes simplex virus gH being gHt and full length herpes simplex virus gL, wherein the gHt and the gL are expressed in and secreted from the cell.
In one embodiment, the gHt comprises herpes simplex virus type 1 gH comprising amino acid residues selected from the group consisting of 1-792, 1-648, 1-475, 1-324 and 1-275.
The invention further relates to an isolated DNA comprising DNA encoding a herpes simplex virus gHt and a substantially full length herpes simplex virus gL.